Rili, Lydia (2024) Exploring Natural Product Biosynthesis in Photorhabdus laumondii: A Novel Strategy through Activation of Bacterial Enhancer Binding Proteins. Masters thesis, Concordia University.
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Abstract
It has been reported that the number of known natural products is much less than the number of biosynthetic gene clusters (BGCs) found in the genomes of microorganisms, which suggests a considerable potential for new drug discovery. However, a significant challenge arises as these BGCs cannot be induced in a laboratory setting. Due to high fitness cost for bacteria, these encoded natural products are tightly regulated. Consequently, our limited understanding of the triggers and regulatory mechanisms of these BGCs impedes progress in the search for potential antibiotics.
To tackle this issue, we propose an alternative approach distinct from the traditional methods such as high-throughput screening, heterologous expression, or the introduction of constitutive promoters. Our strategy involves using the bacterial machinery to activate the expression of BGCs by constitutively activating their bacterial enhancer binding proteins (bEBPs). This is achieved by deleting the regulatory domain located at the N-terminal of bEBPs. The majority of these bEBPs are responsible for activating the σ54-RNA polymerase holoenzyme, thereby initiating transcription.
We have demonstrated σ54’s importance in regulating the expression of a wide range of natural products in Photorhabdus laumondii. This was achieved by utilizing the core domain of DctD (DctD(141-394)), a bEBP from Sinorhizobium meliloti. Further examination of the modified P. laumondi bEBP (mEBPs) revealed diverse effects on natural product expression involving activation and/or repression. Some of these mEBPs exhibit elevated expression levels and, in certain instances, demonstrate propensity for specific natural products compared to DctD(141-394). These preliminary findings suggest that mEBPs, could in time enable us to selectively express a subset of σ54-dependent BGCs. This would bypass the need for specific growth conditions, unlocking a reservoir of previously inaccessible natural products.
| Divisions: | Concordia University > Faculty of Arts and Science > Chemistry and Biochemistry |
|---|---|
| Item Type: | Thesis (Masters) |
| Authors: | Rili, Lydia |
| Institution: | Concordia University |
| Degree Name: | M. Sc. |
| Program: | Chemistry |
| Date: | 1 April 2024 |
| Thesis Supervisor(s): | Findlay, Brandon |
| Keywords: | Natural products (NPs), Photorhabdus laumondii TTO1, bacteria, bacterial enhancer binding proteins (bEBPs), DctD, σ54, biosynthetic gene clusters (BGCs). |
| ID Code: | 993732 |
| Deposited By: | LYDIA RILI |
| Deposited On: | 04 Jun 2024 14:48 |
| Last Modified: | 04 Jun 2024 14:48 |
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